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An increased expression of focal adhesion kinase (FAK) in a variety of cancers is associated with a poor disease prognosis. To study the role of FAK in breast tumor growth and metastasis formation, we used conditional doxycycline-regulated expression of a dominant-negative acting splice variant of FAK, FAK-related non-kinase (FRNK), in MTLn3 mammary adenocarcinoma cells in a syngeneic Fischer 344 rat tumor and metastasis model. In cell culture, doxycycline-mediated expression of FRNK inhibited MTLn3 cell spreading and migration in association with reduced formation of focal adhesions and phosphorylation of FAK on Tyr 397 , but FRNK did not cause apoptosis. Continuous expression of FRNK decreased the primary tumor growth in the mammary fat pad by 60%, which was not due to induction of apoptosis. Lung metastasis formation was almost completely prevented when FRNK was already expressed 1 day before tumor cell injection, whereas expression of FRNK 11 days after injection did not affect lung metastasis formation. FRNK expression during the first 5 days was sufficient to block metastasis formation, excluding the possibility of FRNK-induced dormancy of tumor cells. Together, these data fit with a model wherein FAK is required for breast tumor cell invasion/migration processes that take place in the early phase of metastasis formation. Our findings suggest that FAK is a good candidate for therapeutic intervention of metastasis formation. (Cancer Res 2005; 65(11): 4698-706)

Background: Sublingual allergen immunotherapy (SLIT) has been demonstrated to be both clinically efficacious and safe. However, in line with the current regulatory guidance from the European Medicines Agency, allergen immunotherapy (AIT) products must demonstrate their efficacy and safety in pivotal phase III trials for registration. Objective: We sought to investigate the efficacy and safety of sublingual high-dose liquid birch pollen extract (40,000 allergy units native [AUN]/mL) in adults with birch pollen allergy. Methods: A randomized, double-blind, placebo-controlled, parallel-group multicenter trial was conducted in 406 adult patients with moderate-to-severe birch pollen-induced allergic rhinoconjunctivitis with or without mild-to-moderate controlled asthma. Treatment was started 3 to 6 months before the birch pollen season and continued during the season in 40 clinical study centers in 5 European countries. For primary end point assessment, the recommended combined symptom and medication score of the European Academy of Allergy and Clinical Immunology was used. Secondary end points included quality-of-life assessments, immunologic parameters, and safety.Results: Primary efficacy results demonstrated a significant (P < .0001) and clinically relevant (32%) reduction in the combined symptom and medication score compared with placebo after 3 to 6 months of SLIT. Significantly better rhinoconjunctivitis quality-of-life scores (P < .0001) and the patient's own overall assessment of his or her health status, including the visual analog scale score (Euro Quality of Life Visual Analogue Scale; P 5 .0025), were also demonstrated. In total, a good safety profile of SLIT was observed. Conclusion: This study confirmed both the clinical efficacy and safety of a sublingual liquid birch pollen extract in adults with birch pollen allergy in a pivotal phase III trial (EudraCT: ; g ALIAN s.r.o. Ambulancia alergol ogie a klinickej imunol ogie, Bardejov; and h HAL Allergy B.V., Leiden. The results of this study were presented in part as poster presentations at annual congresses of the European Academy of Allergy and Clinical Immunology (EAACI) in Vienna, Austria, June 2016. Sponsored by HAL Allergy B.V., Leiden, The Netherlands. The sponsor also paid for open access of the article (online open publication). Disclosure of potential conflict of interest: O. Pfaar reports grants and personal fees from HAL Allergy B.V. during the conduct of the study; grants and personal fees from ALK-Abell o, Allergopharma, Stallergenes Greer, HAL Allergy BA.; grants and personal fees from Laboratorios LETI/LETI Pharma; personal fees from Novartis Pharma and MEDA Pharma; grants and personal fees from Anergis S.A.; personal fees from Mobile Chamber Experts (a GA 2 LEN Partner), Pohl-Boskamp, and Indoor Biotechnologies; and grants from GlaxoSmithKline all outside the submitted work. C. Bachert reports grants and personal fees from HAL, ALK-Abell o, Stallergenes Greer, Biotech Tools, Novartis Pharma, Mylan, GlaxoSmithKline, Sanofi Aventis, and Ast...

Focal adhesion kinase (FAK) is up-regulated in a variety of cancers, including breast cancer, in association with poor disease prognosis. In the present study, we examined the role of FAK in the control of anticancer drug-induced apoptosis of mammary adenocarcinoma MTLn3 cells. Doxorubicin caused the formation of well defined focal adhesions and stress fibers early after treatment, which was later followed by their loss in association with the onset of apoptosis. Phosphorylation of FAK on tyrosine 397 decreased only during the onset of doxorubicin-induced apoptosis in a Bcl-2 and caspase-independent manner. Doxorubicin also caused an early activation of protein kinase B (PKB). Expression of the dominant-negative acting focal adhesion kinase-related nonkinase (FRNK) sensitized MTLn3 cells to apoptosis caused by doxorubicin. FRNK inhibited the doxorubicin-induced activation of PKB. In addition, inhibition of phosphatidylinositide-3 (PI-3) kinase with wortmannin inhibited the activation of PKB by doxorubicin. Both wortmannin and transient overexpression of the dual lipid/protein phosphatase and tensin homolog deleted on chromosome 10 enhanced doxorubicin-induced cell death. Altogether, these data fit with a model wherein FAK is involved in the doxorubicin-induced activation of the PI-3 kinase/PKB signaling route, thereby suppressing the onset of apoptosis caused by doxorubicin.Adhesion to the ECM is essential for normal functioning of epithelial cells and provides survival signaling cues (Giancotti and Ruoslahti, 1999;Frisch and Screaton, 2001). In normal cells, detachment generally results in apoptosis, also termed anoikis (Frisch and Francis, 1994;Frisch and Ruoslahti, 1997). However, tumor cells are often resistant to anoikis. This resistance is linked to constitutive activation of survival signaling pathways by oncogenic transformation that otherwise depend on cell adhesion (Streuli and Gilmore, 1999;McFall et al., 2001). These survival signals may confer resistance not only to anoikis but also to apoptosis induced by anticancer drugs due to an overlap in signaling pathways that control cell death. Indeed, adhesion to ECM proteins via the ␤1 integrin subunit protected small cell lung cancer cells against doxorubicin-and etoposide-induced apoptosis and protected breast cancer cells against paclitaxel-induced apoptosis (Sethi et al., 1999;Aoudjit and Vuori, 2001). This dysregulation of cell-ECM interaction-mediated signaling in tumor cells followed by the suppression of or resistance to apoptosis may well contribute to metastasis formation. Likewise, metastatic tumor cells may already be (partly) resistant to anticancer agent-induced apoptosis. Therefore, it is important to know how focal adhesion-dependent signaling is linked to protection against anticancer agent-induced apoptosis. ABBREVIATIONS: ECM, extracellular matrix; AV, Annexin V; CLSM, confocal laser scanning microscopy; FAK, focal adhesion kinase; FRNK, focal adhesion kinase-related nonkinase; GFP, green fluorescent protein; eGFP, enhanced gree...